异基因非清髓造血干细胞移植后WT1特异性T细胞亚群与移植物抗宿主病的关系

Relationship between WT1-specific T-cell subsets and graft-versus-host disease after nonmyeloablative allogeneic transplantation

目的探讨非清髓造血干细胞移植后WT1诱发的各T细胞亚群与移植物抗宿主病（GVHD）之间的关系。方法分析19例表达WTI的白血病患者在全相合非清髓造血干细胞移植后GVHD的发生情况；WT1126—134体外刺激患者外周血单个核细胞，采用胞内因子染色法（ICCS）检测移植前后外周血中Tc1、Tc2、Th1及Th2细胞；对供受者表达HLA—A＊0201的14例患者采用HLA—A＊0201／WTI五聚体检测外周血中的WT1^＋CD8^＋细胞毒性T淋巴细胞（CTL）。结果①19例患者中，17例并发GVHD，其中16例在GVHD发生时出现以Tc1及Th1细胞为主的高峰（P=0．039）。②Ⅱ度以上急性GVHD（aGVHD）患者在GVHD发生时Te1及Th1细胞峰值水平均高于Ⅰ度aGVHD者，但差异无统计学意义（P=0．900及P=0．140）。③弥漫型慢性GVHD（cGVHD）患耆Tc1及Th1细胞峰值水平明显高于局限型cGVHD者，但仅Th1细胞峰值水平差异具有统计学意义（P=0．004），而Tc1细胞峰值水平差异无统计学意义（P=0．060）。④未并发GVHD者Tc1、Th1及WT1^＋CD8^＋CTL峰值水平接近于轻度aGVHD患者的平均水平，但低于中重度aGVHD患者的平均水平。结论GVHD发生时WT1诱发的移植物抗白血病（GVL）效应增强，其强度与GVHD尤其是cGVHD发生的严重程度相关；相对于局限型cGVHD患者，弥漫型cGVHD患者WT1诱发GVL效应的增强可能与Th1细胞有关。

Objective To explore the relationship between WT1-induced T-cell subsets and graftversus-host disease（GVHD） after nonmyeloablative allogeneic hematopoietic stem cell transplantation（NST）. Methods Peripheral blood mononucleated cells （PBMCs） from 19 patients who expressed WT1 and developed GVHD after NST were simulated by WT1126 - 134 peptide in vitro, and proportions of WTl-induced-T- cell subsets （Tc1 ,Tc2 ,Thl ,Th2 Cells） before and after transplant were detected by intracellular cytokine staining （ICCS） assay. WT1-specific CD8 ＋ CTLs of 14 patients with HLA-A ＊ 0201 were detected by HLA-A ＊ 0201/WTI pentamer. Results （1）17 of 19 patients developed GVHD, among whom proportions of Tel and Thl cells, achieved peak value in 16 patients at occurrence of GVHD（ P = 0. 039） ;（2）The peak proportions of Tcl and Thl cells in patients with aGVHD above grade Ⅱ were higher than those with grade Ⅰ , but being no statistical difference （ P = 0. 900 and P = 0. 140, respectively） ; （3）The peak proportion of Thl cells （ P = 0. 004）, but not Tcl cells（P = 0. 060）in patients with extensive cGVHD was significantly higher than that in patients with limited one;（4）Proportions of Tcl, Thl and WT1 ^＋ CD8 ^＋ CTL in patients without GVHD were similar to those in patients with Grade Ⅰ aGVHD, but lower than those in aGVHD above grade Ⅱ. Conclusion GVHD promotes the generation of WTl-induced GVL effect, and the intensity of the latter maybe correlated with the intensity of GVHD, especially cGVHD. Thl cells play a more important role in the enhancement of WT1-induced GVL effect in extensive cGVHD patient than in limited cGVHD patients.